Evidence that DNA polymerase δ contributes to initiating leading strand DNA replication in Saccharomyces cerevisiae

Marta A. Garbacz, Scott A. Lujan, Adam B. Burkholder, Phillip B. Cox, Qiuqin Wu, Zhi-Xiong Zhou, James E. Haber and Thomas A. Kunkel ()
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Marta A. Garbacz: Research Triangle Park
Scott A. Lujan: Research Triangle Park
Adam B. Burkholder: Research Triangle Park
Phillip B. Cox: Research Triangle Park
Qiuqin Wu: Brandeis University
Zhi-Xiong Zhou: Research Triangle Park
James E. Haber: Brandeis University
Thomas A. Kunkel: Research Triangle Park

Nature Communications, 2018, vol. 9, issue 1, 1-11

Abstract: Abstract To investigate nuclear DNA replication enzymology in vivo, we have studied Saccharomyces cerevisiae strains containing a pol2-16 mutation that inactivates the catalytic activities of DNA polymerase ε (Pol ε). Although pol2-16 mutants survive, they present very tiny spore colonies, increased doubling time, larger than normal cells, aberrant nuclei, and rapid acquisition of suppressor mutations. These phenotypes reveal a severe growth defect that is distinct from that of strains that lack only Pol ε proofreading (pol2-4), consistent with the idea that Pol ε is the major leading-strand polymerase used for unstressed DNA replication. Ribonucleotides are incorporated into the pol2-16 genome in patterns consistent with leading-strand replication by Pol δ when Pol ε is absent. More importantly, ribonucleotide distributions at replication origins suggest that in strains encoding all three replicases, Pol δ contributes to initiation of leading-strand replication. We describe two possible models.

Date: 2018
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DOI: 10.1038/s41467-018-03270-4

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